Cell disintegrating apparatus



u 1965 D. FREEDMAN ETAL CELL DISINTEGRATING APPARATUS Filed Oct. 8, 1962 SOUR-CE- 'RECE IVER INVENTORS FEEEDMQN JOHN W. ROSS BY DAV/D ATTQRNE-Y United States Patent 3,1%,56S CELL DlSlNTEGRATING APPARATUS David Freedman, 348 3. 3rd Ave., Highland Park, NJ,

and John W. Ross, 11 Westwood Road, East Brunswick, NJ.

Filed Oct. 9, 1%2, Ser. No. 229,141 7 Claims. (Q1. 241-471) This invention relates to apparatus for disintegrating cells.

There are many types of apparatus known for disintegrating cells. However, such known types of apparatus are not universally useful in the bacteriological arts. Some types of apparatus, for example sonic devices, operate efliciently with only certain types of bacteria. Other types of devices which employ abrasive materials for performing the disintegrating operation are not completely efficient and generate excessive heat which cannot be readily removed. Still other types of apparatus are ineflicient and unable to disintegrate a large number of cells in a short period of time.

Accordingly, the objects of the invention concern the provision of improved apparatus for disintegrating cells, the apparatus being characterized by an optimum combination of efficiency, capacity, and universality.

The apparatus of the invention utilizes high speed shaking with rapid reversals in direction of acceleration to produce quick and efiicient cellular disintegration. Briefly, the apparatus includes a cartridge or container for material to be disintegrated and mechanical means for oscillating or shaking the container at high speed with a short stroke. The entire apparatus is supported and constructed to avoid any loss of energy due to vibration or the like. The cartridge includes an optimum combination of features to provide efficient rupture of the material contained therein by crushing, shearing, grinding, and cavitation and is oriented physically to promote this end. The apparatus may be operated continuously or with separate batches of material.

The invention is described in detail by reference to the drawing wherein:

FIG. 1 is a perspective view of apparatus embodying the invention; and

FIG. 2 is a sectional View of a portion of the apparatus shown in FIG. 1 and a schematic representation of other apparatus associated therewith.

Referring to the drawing, cell disintegrating apparatus 1% embodying the invention is mounted on a vibrationabsorbing base 12 which carries a pad 14 of resilient material, for example rubber, on which a main support plate 15 is mounted. The plate 16 supports the operating portions of the invention which include a drive shaft (not shown per se) mounted in a well-lubricated bearing housing 13 which is rigidly supported on a pair of plates 2E). The plates 20 are welded or otherwise secured to the main support plate 16 and are spaced apart a suitable distance to provide optimum, vibration-free support for the shaft housing 13. The plates 20 include generally semicircular cutout portions 22 which receive the bearing housing. The primary drive shaft within housing 18 is coupled at one end to a suitable variable speed power source such as a motor 23 or the like. The drive shaft carries at the other end an adjustable stroke eccentric 24 which automatically sets the amount of counterbalance with the adjustment of stroke. The eccentric 24 is coupled to, and transmits reciprocating motion to, a drive arm which is rotatably secured thereto.

The drive arm 30 is coupled at its free end 32 to a cartridge-holding arm 44 which is pivotally secured to a rugged and rigid base support 46 by a pin 43. The base 46 is suitably secured to main support plate 16 by welding or the like. The upper end of arm 44 remote from pin 48 is provided with a bore 49 in which is secured a container or cartridge 59 in which the material to be disintegrated is carried. The cartridge is oriented with its long axis horizontal and perpendicular both to the longitudinal axis of arm 44 and to the direction in which arm 44 reciprocates. The cartridge-holding arm 44 has an aperture 56 suitably positioned between its ends which is adapted to receive the arm 30. The free end of arm 30 is inserted in the aperture and is rotatably secured to arm 44 by means of a pin 60.

As shown in FIG. 2, in order to provide secure mounting of the cartridge 50 in the bore 49 in arm 44, the bore is tapered in diameter and a complementarily tapered squeeze sleeve 72 is provided surrounding the cartridge 59. The cartridge is locked in place by means of a squeeze nut 32 which is threaded on its outside diameter to fit the correspondingly threaded portion of bore 49. Other cartridge locking arrangements might be used as desired.

Referring to FIG. 2 for details of construction of the cartridge 59, the cartridge comprises a hollow open-ended tubular housing 86 which is preferably made of metal, for example, stainless steel or aluminum. The inner surface 88 of housing should be as hard as possible, and if the housing is of aluminum, the surface 83 might be provided with a layer of aluminum oxide. The housing 36 may have any suitable cross-section; for example, it may be circular, elliptical, square, or the like.

A metal center bar or bathe W of aluminum with hardened surface or stainless steel or the like is provided within the housing 86 and is positioned on the longitudinal axis of the housing. The bafile bar 99 may be solid or hollow. One end of the center bar 9@ is provided with a hollow tubular extension 96 which comprises an outlet tube for the cartridge. The other end 97 of center bar 90 is provided with a conical aperture or bore 93 at its cen for. The end 94 of bar 90 is provided with an aperture 1%, through which material that has been disintegrated is fed from the inside of the housing 86 into outlet tube 96. Disintegrated material may thus be removed from the housing 86.

The center bar 95 is held in place in the housing by means of the foliowing apparatus. An inlet end cap 104, preferably of metal, is threaded on one end of the housing 86 with an O ring seal 18-6 provided between adjacent abutting portions of the two members. The inlet and cap is provided with a threaded central opening 198, in which a correspondingly threaded hollow inlet tube 112 is mounted. A lock nut 116 is provided for locking the inlet tube in position in the end cap. The inner end of the inlet tube is provided with a conical head Hi2 which is adapted to engage the bore 98 in the adjacent end of the center bar 9%) which is supported thereby. The portion of the inlet tube adjacent to the conical head is provided wtih an inlet aperture 124, through which material to be disintegrated may be fed into the housing.

A similar outlet end cap is provided threaded on the other end of the tubular housing 36 with an O ring seal 134 between the members. The outlet end cap is provided with a threaded axial bore in which the hollow outlet tube portion 96 of the center bar, which is also threaded, is adjustably positioned. A lock nut 144 is provided for locking the outlet tube in place in the end cap. A wire mesh strainer or other suitable strainer (sin tered metal, metal wool, etc.) 148 is mounted adjacent to the inner wall of the outlet end cap and surrounding the end portion 9 of the center bar 9%) and held in place by a collar on bar 99 or by an adjacent larger diameter portion of the center bar or by some other suitable means. The strainer is of a sufliciently fine mesh or porosity so that it allows disintegrated material to pass out of the cartridge but retains within the cartridge a plurality of small diam- 'tents of cartridge 50 during operation.

or eter plastic beads 152. The beads are provided within the cartridge to act as abrasive particles and to olfer a large surface area for impact and shearing of the cell material to be distintegrated. The beads are of low density, preferably a density similar to that of the liquid which carries cellular material. The beads are also of small diameter, in the range of about 0.1 mm. to 1.0 mm. to provide maximum surface area with minimum weight so that minimum energy is absorbed and minimum heat is generated thereby. Beads of styrene-divinylbenzene are suitable.

The inlet tube 112 is coupled through suitable flexible tubing 168 to a source 169 of cells to be disintegrated. The cellular material is provided in a suitable liquid carrier. The cellular material is fed to the" cartridge 50 means of a variable speed pump 172 such as a peristaltic pump or the like, which is adjusted for the desired flow rate. A compressible gas, for example air, is also fed to the cartridge mixed with the cellular material. The outlet pipe 140 is also coupled through flexible tubing 174 to a suitable receiver 176 for the disintegrated cellular .material.

The apparatus of the invention also preferably includes some provision for controlling the temperature of the con- For example, a controlled stream of carbon dioxide may be sprayed on the cartridge as it is shaken, or the cellular material may be suitably precooled in the source 169. If desired, the cartridge 50 may be contained within a controlled environment with forced air or fluid circulation included.

The apparatus of the invention operates on a raw material charge which includes a gas such as air, nitrogen or oxygen, a carrier liquid, and the cellular material to be distintegrated. In charging the cartridge 50, the pump 172 is operated to admit a quantity of raw material to the cartridge, and the cartridge is agitated or oscillated with a stroke of A inch to inch at high speed which is preferably of the order of 6,000 to 8,000 cycles per minute. The orientation of the cartridge, the center bar, the compressible gas phase, and the plastic beads combine to provide highly efiicient cell disintegration. When the raw material has been considered to be properly disintegrated, the contents of the cartridge is removed. Then, the cartridge is charged again, and the operation is repeated. The charging and withdrawal operations can be performed while the cartridge is being agitated. This type of operation is known as batch feeding. The apparatus may also be operated continuously, with material being fed steadily into the cartridge as it is reciprocated with the speed of material feed and reciprocation being adjusted for the type of material being processed.

The apparatus of the invention has been found to disintegrate quickly and efiiciently, and without the generation of excessive heat, a wide variety of cellular materials including bacteria, molds, actinomycetes and yeasts. The apparatus is also able to process a quantity of material in amounts ranging from about 800 milliliters per hour to about 5 liters per hour, depending on the material. Subcellular preparations also retain a high level of enzymatic activity due to the speed and efiiciency of the machine.

In normal operation, the apparatus of the invention has been used to rupture cell suspensions as follows where the concentration is given in terms of mg. of cell nitrogen per ml.: 12 mg. per ml. of Aerobacter aerogenes in about 96 seconds; 9 mg. per ml. of Bacterium cyclooxydans in about 35 seconds; 4 mg. per ml. of Bordetella pertussis in about 45 seconds; 3 mg. per ml. of Escherichia coli in about 85 seconds; 3.5 mg. per ml. of Pencillium chrysogenum in less than 20 seconds; 15.5 mg. per ml. of Sacclzaromyces cerevisae in less than 17 seconds; and 7.5 mg. per ml. of Streptomyces nodosus in about 85 seconds.

There is no other commercially available apparatus which can rupture completely all of these types or comparable quantities of cells within a reasonable time, and, in addition, apparatus which can rupture selected types of cells are considerably less efiicient. For example, according to published data, a sonic device which carries the name Sonifier can rupture completely bacteria in 1 to 11 minutes, yeast in 5 to 50 minutes, molds in 3 minutes, and actinomycetes in 1 minute. A device known by the name Biosonic can rupture Escherichia coli in about 10 minutes. A known type of pressure device provides 70% to rupture of bacteria and yeast with with two to three passes through an orifice, with each cycle taking several minutes. Finally, a known colloid mill requires more than 5 minutes to rupture Escherichia coli and more than 10 minutes to rupture Bakers yeast.

The theory of operation of the apparatus of the invention is that shaking of the chamber contents, including a compressible gas phase, causes violent agitation which results in cavitation, crushing, shearing, and grinding action and impact of the cells on each other, on the cartridge wall, the center bar, and the plastic beads, and that such action causes disintegration of the cells. These eifects can be amplified by increasing the frequency of oscillation and the stroke of the cartridge.

What is claimed is:

1. Cell disintegrating apparatus comprising a vibration-absorbing base,

a support member secured to said base,

a cartridge-carrying arm pivotally secured at one end to said support member and adapted to be oscillated in a predetermined direction,

the other end of said arm having an aperture and a cartridge mounted in said aperture,

said cartridge having a longitudinal axis oriented horizontally and perpendicular to the direction in which said housing is adapted to oscillate,

abrasive means inside said cartridge for disintegrating organic material provided therein,

means coupled to said cartridge for feeding material thereto and for withdrawing material therefrom, and

drive means coupled to said arm for oscillating said housing at high speed in said predetermined direction whereby the material contained in said cartridge is disintegrated.

2. The apparatus defined in claim 1 wherein said arm is generally vertically oriented and has a lower end secured to said support member and an upper end provided with said aperture for receiving said cartridge, said drive means being secured to said arm between its ends.

3. The apparatus defined in claim 1 wherein said arm is generally vertically oriented and has a lower end secured to said support member and an upper end provided with said aperture for receiving said cartridge, said aperture being tapered and having :a correspondingly tapered sleeve surrounding said container, said drive means being secured to said arm between its ends.

4. The apparatus defined in claim 1 wherein said arm is generally vertically oriented and has a lower end secured to said support member and an upper end provided with said aperture for receiving said cartridge, said drive means being secured to said arm between its ends, said drive means including an adjustable eccentric for adjusting the stroke of oscillation of said arm.

5. The apparatus defined in claim 1 wherein said cartridge comprises a metal cylinder and contains an axial metallic bar extending along its length and a plurality of small diameter, low density beads.

6. The apparatus defined in claim 1 wherein said cartridge comprises a metal cylinder and contains an axial metallic bar extending along its length and a plurality of small diameter, low density beads of a synthetic resinous material.

7. The apparatus defined in claim 1 wherein said cartridge comprises a metal cylinder and contains an axial metallic bar extending along its length and a plurality of small diameter, low density beads of a synthetic resinous material,

and fluid conduits coupled to the ends of said metal cylinder and adapted one to admit cellular material to the cartridge and one to withdraw cellular material from said cartridge, a fluid-tight seal being provided at the ends of said cylinder with said fluid conduits.

References fitted by the Examiner UNITED STATES PATENTS 2,168,093 8/39 Osgood 241170 X 6 2,315,229 3/43 Schieferstein 241-175 2,468,515 4/49 Robinson 241170 FOREIGN PATENTS 5 240,022 8/ 62 Australia.

OTHER REFERENCES Mudd et al.: A Low Temperature Ball Mill for the Liberation of Labile Cellular Products. Public Health 10 Reports, vol 52, No. 27, July 2, 1937, pages 167-78.

(Copy in Group 340 in 241Hand C Digest.)

J. SPENCER OVERHOLSER, Primary Examiner. 

1. CELL DISINTEGRATING APPARATUS COMPRISING A VIBRATION-ABSORBING BASE, A SUPPORT MEMBER SECURED TO SAID BASE, A CARTRIDGE-CARRYING ARM PIVOTALLY SECURED AT ONE END TO SAID SUPPORT MEMBER AND ADAPTED TO BE OSCILLATED IN A PREDETERMINED DIRECTION, THE OTHER END OF SAID ARM HAVING AN APERTURE AND A CARTRIDGE MOUNTED IN SAID APERTURE, SAID CARTRIDGE HAVING A LONGITUDINAL AXIS ORIENTED HORIZONTALLY AND PERPENDICULAR TO THE DIRECTION IN WHICH SAID HOUSING IS ADAPTED TO OSCILLATE, ABRASIVE MEANS INSIDE SAID CARTRIDGE FOR DISINTEGRATING ORGANIC MATERIAL PROVIDED THEREIN, MEANS COUPLED TO SAID CARTRIDGE FOR FEEDING MATERIAL THERETO AND FOR WITHDRAWING MATERIAL THEREFROM, AND DRIVE MEANS COUPLED TO SAID ARMS FOR OSCILLATING SAID HOUSING AT HIGH SPEED IN SAID PREDETERMINED DIRECTION WHEREBY THE MATERIAL CONTAINED IN SAID CARTRIDGE IS DISINTEGRATED. 